Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
one-generation reproductive toxicity
Remarks:
based on test guideline (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
3 August 1987 to 12 January 1988
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
A GLP study conducted to sound scientific principles, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. As the study was conducted with the structural analogue, bis(2-ethylhexyl) adipate, it has been assigned a reliability score of 2.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 28 days
- Weight at study initiation: (P) Males: 72.5 g; Females: 71.1 g
- Housing: 2 females or 1 male per cage
- Water (e.g. ad libitum): filtered tap water
- Acclimatisation period: 6-7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 21
- Humidity (%): 45-60
- Air changes (per hr): 15 - 25
- Photoperiod (hrs dark / hrs light): 12 /12

IN-LIFE DATES: From: 3 August 1987 To: 12 January 1988
Route of administration:
oral: feed
Vehicle:
other: hexane
Details on exposure:
DIET PREPARATION
- Mixing appropriate amounts with (Type of food):
Dose level 300 ppm: 9.07g/30 kg
Dose level 1800 ppm: 54.44g/30 kg
Dose level 12000 ppm: 362.90g/30 kg
Details on mating procedure:
- M/F ratio per cage: 1 male and 2 female per cage
- Length of cohabitation: 10 days
- Proof of pregnancy: vaginal smear were examined daily
- After 10 days of unsuccessful pairing replacement of first male by another male with proven fertility
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): separately
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Mean concentrations within 2 % of target concentration for all groups. Test material was not detected in any control diet (detection limit 10 ppm). Chemical stability of test material in diet was determined on three batches of diet at nominally 300 ppm and 12000 ppm. Satisfactory chemical stability was established.
Homogeneity of test material in diet mixtures was satisfactorily demonstraded on the first diet batch at nominally 300 and 12000 ppm test material.
Duration of treatment / exposure:
10 weeks
Frequency of treatment:
The rats in each generation were fed experimental diets continuously until termination.
Remarks:
Doses / Concentrations:
0, 300, 1800, 12000 ppm
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
0, 28, 170, 1080 mg/kg
Basis:
nominal in diet
No. of animals per sex per dose:
30 females and 15 males per group in total 4 groups.
Control animals:
yes, plain diet
Details on study design:
According to randomisation the rats where housed
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes, at least once daily

DETAILED CLINICAL OBSERVATIONS: Yes, once weekly

BODY WEIGHT: Yes, of all rats were recored at weekly intervals throughout the premating period.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, each cage of rats was recorded throughout the premating periods and calculated on a weekly basis. The food utilisation value per cage was calculated as the weight gained by the animals in the cage per 100g of food eaten.
Oestrous cyclicity (parental animals):
no data
Sperm parameters (parental animals):
no data
Litter observations:
STANDARDISATION OF LITTERS
A count of all live and dead pups was made within 24 hrs (day 1), at days 5, 11, 22, 29 and 36 post partum. The sexes of the pups were also recorded at these times.
Postmortem examinations (parental animals):
SACRIFICE
All animals at scheduled kills and those killed during the study were anaesthetised by inhalation of halothane BP vapour and killed by exsanguination. All surviving males were killed after completion of mating. All females were killed after weaning thier litters.

Histological examination: cervix, epididymis, liver, mammary gland, ovary, prostate, seminal vesicle, testis, uterus, abnormal tissues.
Postmortem examinations (offspring):
All pups were killed as soon as possible after Day 36 post partum.

Histological examination: cervix, epididymis, liver, mammary gland, ovary, prostate, seminal vesicle, testis, uterus, abnormal tissues.
Statistics:
Mean bodyweight gain, food consumption and food utilisation during the premating period, female bodyweight gain during pregnancy, parental liver weights and pup (litter) bodyweight gain until Day 36 post partum.
Reproductive indices:
Mean length of gestation, mean pre-coital interval
Offspring viability indices:
Mean live born index, mean survival index, mean litter size, total litter weight and whole litter losses.
Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
See "Details on results (parental animals)" for information
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
See "Details on results (parental animals)" for information
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Test substance intake: See "Details on results (parental animals)" for information
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
TEST SUBSTANCE INTAKE
There was a slight increase in food consumption in males dosed with 12000ppm test material from 6-10 weeks of the study, the effect being statistically significant at weeks 6-9. Food utilisation was slightly less efficient overall for males receiving 12000ppm test material.

ORGAN WEIGHTS
An increase in liver weight was observed for both male and female parents receiving 12000ppm test material. No other group treatment group was effected. This increase in liver weight has been reported previously and is associated with peroxisome proliferation (Moody and Reddy 1978).
Dose descriptor:
NOAEL
Effect level:
170 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: increased absolute liver weights, and reduced body weight gain in females
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
BODY WEIGHT
Mean pup weight gain and total litter weight for both male and female offspring receiving 12000 ppm test material were reduced throughout the whole of the post partum phase. There was no effect on either male or female pup weight gain in any other dose group in comparison with the control animals.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
170 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Reproductive effects observed:
not specified
Conclusions:
Under the conditions of the study the No Observed Adverse Effect Level for parental animals was determined to be 170 mg/kg bw/day. The No Observed Adverse Effect Level for the F1 offspring was determined to be 170 mg/kg bw/day.
Executive summary:

The reproductive toxicity of the test material was determined in a GLP study conducted to a methodology which was similar or equivalent to that which is outlined in the standardised guideline OECD 415. During the study groups of 30 females and 15 males received test material in the diet at dose levels of 0, 300, 1800, 12000 ppm in diet. Animals were administered test material daily over a period of 10 weeks. One male rat was cohabited with two female rats, for 10 days, until mating was confirmed. Animals were caged separately thereafter. During the study, animals were observed daily for clinical signs and body weights were recorded at weekly intervals. Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day and food utilisation values per cage were calculated (as the weight gained by the animals in the cage per 100g of food eaten). A count of all live and dead pups was made within 24 hrs (day 1), at days 5, 11, 22, 29 and 36 post partum. The sexes of the pups were also recorded at these times. Parental males were sacrificed after completion of mating and parental females were sacrificed after weaning their litters for histopathological examination. All pups were killed as soon as possible after Day 36 post partum and subjected to histopathological examination.

In parental animals, there was a slight increase in food consumption in males dosed with 12000 ppm test material from 6 - 10 weeks of the study, the effect being statistically significant at weeks 6 - 9. Food utilisation was slightly less efficient overall for males receiving 12000 ppm test material. An increase in liver weight was observed for both male and female parents receiving 12000ppm test material. No other group treatment group was effected.

In F1 offspring, Mean pup weight gain and total litter weight for both male and female offspring receiving 12000 ppm test material were reduced throughout the whole of the post partum phase. There was no effect on either male or female pup weight gain in any other dose group in comparison with the control animals.

Under the conditions of the study the No Observed Adverse Effect Level for parental animals was determined to be 170 mg/kg bw/day. The No Observed Adverse Effect Level for the F1 offspring was determined to be 170 mg/kg bw/day.

Effect on fertility: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
170 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The study was conducted to GLP, and sound scientific principles, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. As the study was conducted with the structural analogue, bis(2-ethylhexyl) adipate, it has been assigned a reliability score of 2.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

The reproductive toxicity of the test material was determined in a GLP study conducted to a methodology which was similar or equivalent to that which is outlined in the standardised guideline OECD 415. During the study groups of 30 females and 15 males received test material in the diet at dose levels of 0, 300, 1800, 12000 ppm in diet. Animals were administered test material daily over a period of 10 weeks. One male rat was cohabited with two female rats, for 10 days, until mating was confirmed. Animals were caged separately thereafter. During the study, animals were observed daily for clinical signs and body weights were recorded at weekly intervals. Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day and food utilisation values per cage were calculated (as the weight gained by the animals in the cage per 100g of food eaten). A count of all live and dead pups was made within 24 hrs (day 1), at days 5, 11, 22, 29 and 36 post partum. The sexes of the pups were also recorded at these times. Parental males were sacrificed after completion of mating and parental females were sacrificed after weaning their litters for histopathological examination. All pups were killed as soon as possible after Day 36 post partum and subjected to histopathological examination.

In parental animals, there was a slight increase in food consumption in males dosed with 12000 ppm test material from 6 - 10 weeks of the study, the effect being statistically significant at weeks 6 - 9. Food utilisation was slightly less efficient overall for males receiving 12000 ppm test material. An increase in liver weight was observed for both male and female parents receiving 12000ppm test material. No other group treatment group was effected.

In F1 offspring, Mean pup weight gain and total litter weight for both male and female offspring receiving 12000 ppm test material were reduced throughout the whole of the post partum phase. There was no effect on either male or female pup weight gain in any other dose group in comparison with the control animals.

Under the conditions of the study the No Observed Adverse Effect Level for parental animals was determined to be 170 mg/kg bw/day. The No Observed Adverse Effect Level for the F1 offspring was determined to be 170 mg/kg bw/day.

The existing data are considered adequate to address this endpoint and a two-generation reproduction toxicity study is considered scientifically unnecessary.


Short description of key information:
NOAEL (parental) = 170 mg/kg/day, NOAEL (offspring) = 170 mg/kg bw/day, male/female rat, Cefic 1988b

Justification for selection of Effect on fertility via oral route:
Only one study is available.

Effects on developmental toxicity

Description of key information
NOAEL (maternal toxicity)  = 170 mg/kg/day, NOEL (fetotoxicity) = 28 mg/kg bw/day, rat, Cefic 1988b
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 September 1987 to 16 October 1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
A GLP study conducted to sound scientific principles, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. As the study was conducted with the structural analogue, bis(2-ethylhexyl) adipate, it has been assigned a reliability score of 2.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
Principles of method if other than guideline:
For determination of limit dose the OECD 414 guideline was followed. The bottom dose level was selected based on information obtained from literature and was related to likely human exposure.
The maximum human intake has been estimated by MAFF (UK) 1986 to be 16 mg/day and this was calculated to be 25 mg/kg/day for a 60-70 kg human. A factor of 100 was then used to provide an appropriate margin of safety which thus gave a dose of 25 mg/kg/day in rats for the present study. The middle dose was spaced between these two doses using approximately a six-fold factor . The dose levels were then calculated
as ppm in the diet (for a 300g rat eating 25g food per day). The rats were dosed on Days 1-22 inclusive of gestation, Day 1 being the day that mating was confirmed by a sperm-positive vaginal smear.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS:
- Strain: Wistar rats of the Alpk: AP fSD strain
- Age at study initiation: aproximately 12 weeks
- Weight at study initiation: 218 - 278 g
- Housing: individually housed in rat racks supplied by All Type Tools Ltd, Woolwich, London, UK
- Water: tap water ad libitum via an automatic dispensary system

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 24 °C
- Humidity (%): 44 - 70 %
- Air changes (per hr): 12 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light

IN-LIFE DATES: From 15 September 1987 to 16 October 1987
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION
The experimental diets were prepared in 30kg batches from premixes and dispensed into glass feeding jars . Two batches of diet were prepared at each level.

Diet sampling and analysis:
A sample was taken from each diet prepared . Samples were taken from the diet feeding jars and analysed. Chemical stability of DEHA in CT1 diet was determined at 300 and 12000ppm .
Homogeneity of DEHA was also examined in a concurrent study (Tinston 1988) and found to be satisfactory
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The amount of food consumed by each animal was measured daily by giving a weighed quantity of food contained in a glass jar on one day and calculating the amount consumed from the residue on the next .
Details on mating procedure:
Wistar-derived, virgin female rats were paired overnight at the Breeding Unit with unrelated males of the same strain . On the following morning, vaginal smears from these females were examined for the presence of sperm. The day when spermatozoa were detected was designated Day 1 of gestation and on this same day, successfully mated females were delivered to the experimental unit at CTL .
A total of 96 mated females was supplied over a two week period . Twelve female rats were supplied on each of eight days .
Duration of treatment / exposure:
From Day 1 of gestation until termination on Day 22.
Frequency of treatment:
Daily
Duration of test:
All females were sacrificed on Day 22 of gestation
Remarks:
Doses / Concentrations:
0, 300, 1800, 12000 ppm
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
0, 28, 170, 1080 mg/kg
Basis:
nominal in diet
No. of animals per sex per dose:
24 females per dose group
Control animals:
yes, plain diet
Details on study design:
The study was divided into 24 replicates (randomised blocks) with each replicate containing one rat from each dosage group. Cages within the
replicates were assigned to one of the four groups using computer-generated random number permutations. The individual animal numbers were then assigned sequentially within the relevant groups to give the rack plan. On arrival (Day 1 of gestation) each rat was allocated to a cage (and therefore a treatment group) randomly within the replicate and individually identified by ear punching with the number assigned to it from the experimental design. Replicates were filled sequentially with three replicates added to the study on each of the eight days on which rats were received.
Maternal examinations:
Clinical observations:
All animals were checked on arrival to ensure that they were physically normal externally. They were subsequently observed daily for any changes in behaviour or clinical condition and these were recorded.

Body weight:
The bodyweight of each animal was recorded daily on Days 1 to 22 inclusive of gestation.

Terminal Investigations:
On Day 22 of gestation all the animals were killed by over exposure to halothane BP (FLUOTHANE, ICI Pharmaceuticals, Macclesfield, Cheshire, UK) vapour. A post mortem was performed and all animals were examined macroscopically. The intact gravid uterus (minus ovaries and trimmed free of connective tissue) was removed and weighed.
Ovaries and uterine content:
The ovaries and uterine content were examined after termination. The following data was recorded:
Number of corpora lutea in each ovary.

Number and position of implantations subdivided into:
(a) live foetuses
(b) early intra-uterine deaths
(c) late intra-uterine deaths

Intra-uterine deaths were classified as follows:
Early intra-uterine deaths showed decidual or placental tissue only. Late intra-uterine deaths showed embryonic or foetal tissue in addition to placental tissue. The implantations were assigned letters of the alphabet to identify their position in utero starting at the ovarian end of the left horn and ending at the ovarian end of the right horn.
Fetal examinations:
Each foetus was weighed and individually identified within the litter by means of a cardboard tag. After weighing, the foetuses were killed with an intra-cardiac injection of pentobarbitone, sodium solution, 200 mg/ml, (EUTHATAL, May and Baker Ltd, Dagenham, Essex, UK).

Assessment of Teratogenicity:
Each foetus was examined for external abnormalities and for cleft palate. All foetuses were then examined internally for visceral abnormalities under magnification, sexed, eviscerated and fixed in methanol. The head of each foetus was cut along the fronto-parietal suture line and the brain was examined for macroscopic abnormalities. (The brains of one litter, female 72, 1800ppm, inadvertently were not examined). The carcasses were then returned to methanol for subsequent processing and staining with Alizarin Red S. The stained foetal skeletons were examined for abnormalities and the degree of ossification was assessed. The individual bones of the manus and pes were assessed and the result converted to a four point scale. Abnormalities were classified as major (rare or possibly lethal or both) or minor (deviations from normal that are not uncommon at external, visceral or skeletal examination) defects. Variations were also recorded and classified as minor defects or variants depending on the historical frequency of occurrence in rats of this strain.
Statistics:
The following data were considered by analysis of variance:
(i) Maternal bodyweight gain
(ii) Maternal food consumption
(iii) The numbers of implantations and live foetuses per female
(iv) Percentage pre-implantation loss and percentage post-implantation loss (calculated on an individual litter basis). The percentage pre-implantation loss and post-implantation loss were transformed before analysis using the double arcsine transformation of Freeman and Tukey (1950). The analyses of variances were weighted by the denominator in the proportion.
(v) The percentage of implantations which were early intra-uterine deaths (calculated on an individual litter basis).
(vi) Gravid uterus weight, litter weight and mean foetal weight (calculated on an individual litter basis).
(vii) Mean manus and pes score per foetus (calculated on an individual litter basis).
(viii) The percentage of foetuses with minor external/visceral defects only, external/visceral variants and minor skeletal defects only (calculated on an individual litter basis).

The analyses of variance allowed for the replicate structure of the study design and were carried out using the GLM procedure in SAS (1985). Unbiased estimates of the treatment group means were provided by the least square means (LSMEANS option in SAS). Individual treatment group means were compared with the control group mean using Student's t-test based on the error mean square in the analysis.

Further analysis:
Fisher's Exact Test, comparing each treated group with the control group.
All statistical tests were one-sided with the following exceptions which were two-sided: maternal bodyweight gain, maternal food consumption and the proportion of male foetuses.
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
Administration of 12000 ppm test material resulted in slight maternal toxicity (small maternal reduction in body weight gain and reduced food
consumption). At 1800 ppm test material, there was no evidence of maternal toxicity.
Dose descriptor:
NOAEL
Effect level:
170 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
Administration of 1800 ppm test material resulted in minimal foetotoxicity (reduced ossification and increase in the incidence of visceral variants). A dietary level of 300 ppm test material was a clear no-effect level for embryonic development.
Dose descriptor:
NOEL
Effect level:
28 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: fetotoxicity
Abnormalities:
not specified
Developmental effects observed:
not specified

Diet analysis: chemical stability of test material in diet was satisfactory. Concentrations of test material were within acceptable limits.

 

Maternal clinical observations: All rats survived to scheduled termination.

The clinical findings observed were considered not to be related to test material administration.

 

Maternal body weight: Administration of 12000 ppm test material was associated with a small but statistically significant reduction in bodyweight gain compared with the control group which was most marked at the start of the feeding period.

 

Maternal food consumption: Maternal food consumption was statistically significantly reduced in the 12000ppm group from Days 2-18 inclusive of pregnancy. There were no adverse effects on food consumption in the 300 or 1800 ppm test material groups.

 

Maternal macroscopic findings (post mortem): macroscopic changes were considered not to be related to test material treatment. There was no effect at any dose on foetal weight, litter weight, gravid uterus weight, numbers of intra-uterine deaths or numbers of external abnormalities.

Foetal examinations: The incidence of minor external and visceral defects was unaffected by treatment although two visceral variants were increased at the top two dose levels; kinked ureter being increased in the 1800 and 12000 ppm groups and slightly dilated ureter being increased in the 12000 ppm group. Overall, minor skeletal defects were increased in a dose-related manner at 1800 and 12000 ppm test material, while skeletal variants and pes score were increased at the top dose only. These findings indicate slightly poorer ossification at the 1800 and 12000 ppm dose levels. The reduced ossification and increase in the incidence of visceral variants are considered to be the result of slight foetotoxicity. There was no treatment-related effect on skeletal or visceral variants at 300 ppm test material.

Conclusions:
Under the conditions of the study the maternal toxicity No Observed Adverse Effect Level was determined to be 170 mg/kg bw/day while the developmental toxicity No Observed Effect Level was determined to be 28 mg/kg bw/day.
Executive summary:

The developmental toxicity of the test material was determined in a GLP study which was conducted to a methodology similar to that which is outlined in the standardised guideline OECD 414. During the study 24 females were mated with unrelated males and received test material in the diet, at dose levels of 0, 300, 1800, 12000 ppm, from day 1 to day 22 of gestation. During treatment females were observed daily for clinical signs and individual body weights were recorded. On day 22 of gestation all females were sacrificed and examined macroscopically.

 

The intact gravid uterus (minus ovaries and trimmed free of connective tissue) was removed and weighed. The ovaries and uterine content were examined after termination and the number of corpora lutea and number and position of implantations were recorded. After weighing, each foetus was sacrificed and examined for external abnormalities and for cleft palate. All foetuses were then examined internally for visceral abnormalities under magnification, sexed, eviscerated and fixed in methanol. The head of each foetus was examined for macroscopic abnormalities. The foetal skeletons were examined for abnormalities and the degree of ossification was assessed. The individual bones of the manus and pes were assessed and the result converted to a four point scale. Abnormalities were classified as major (rare or possibly lethal or both) or minor (deviations from normal that are not uncommon at external, visceral or skeletal examination) defects. Variations were also recorded and classified as minor defects or variants depending on the historical frequency of occurrence in rats of this strain.

 

All maternal females survived until scheduled sacrifice and no adverse clinical signs of toxicity were observed. Maternal food consumption was statistically significantly reduced in the 12000 ppm group from Days 2-18 inclusive of pregnancy. There were no adverse effects on food consumption in the 300 or 1800 ppm test material group. In line with the reduction in food consumption in the high-dose females, administration of 12000 ppm test material was associated with a small but statistically significant reduction in bodyweight gain compared with the control group which was most marked at the start of the feeding period. Maternal macroscopic findings (post mortem) were considered not to be related to test material treatment. There was no effect at any dose on foetal weight, litter weight, gravid uterus weight, numbers of intra-uterine deaths or numbers of external abnormalities.

 

Examination of the feotuses revealed that the incidence of minor external and visceral defects was unaffected by treatment although two visceral variants were increased at the top two dose levels; kinked ureter being increased in the 1800 and 12000 ppm groups and slightly dilated ureter being increased in the 12000 ppm group. Overall, minor skeletal defects were increased in a dose-related manner at 1800 and 12000 ppm test material, while skeletal variants and pes score were increased at the top dose only. These findings indicate slightly poorer ossification at the 1800 and 12000 ppm dose levels. The reduced ossification and increase in the incidence of visceral variants are considered to be the result of slight foetotoxicity. There was no treatment-related effect on skeletal or visceral variants at 300 ppm test material.

 

Overall, under the conditions of the study, the NOAEL (maternal toxicity), based on reduction in body weight and food intake at 12000 ppm (1080 mg/kg), was determined to be 170 mg/kg bw/day while the NOEL (fetotoxicity), based on reduced ossification and increase in the incidence of visceral variants observed at the highest doses, was determined to be 28 mg/kg bw/day.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Study duration:
subacute
Species:
rat
Quality of whole database:
The study was conducted to GLP, and sound scientific principles, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. As the study was conducted with the structural analogue, bis(2-ethylhexyl) adipate, it has been assigned a reliability score of 2.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

The developmental toxicity of the test material was determined in a GLP study which was conducted to a methodology similar to that which is outlined in the standardised guideline OECD 414. During the study 24 females were mated with unrelated males and received test material in the diet, at dose levels of 0, 300, 1800, 12000 ppm, from day 1 to day 22 of gestation. During treatment females were observed daily for clinical signs and individual body weights were recorded. On day 22 of gestation all females were sacrificed and examined macroscopically.

 

The intact gravid uterus (minus ovaries and trimmed free of connective tissue) was removed and weighed. The ovaries and uterine content were examined after termination and the number of corpora lutea and number and position of implantations were recorded. After weighing, each foetus was sacrificed and examined for external abnormalities and for cleft palate. All foetuses were then examined internally for visceral abnormalities under magnification, sexed, eviscerated and fixed in methanol. The head of each foetus was examined for macroscopic abnormalities. The foetal skeletons were examined for abnormalities and the degree of ossification was assessed. The individual bones of the manus and pes were assessed and the result converted to a four point scale. Abnormalities were classified as major (rare or possibly lethal or both) or minor (deviations from normal that are not uncommon at external, visceral or skeletal examination) defects. Variations were also recorded and classified as minor defects or variants depending on the historical frequency of occurrence in rats of this strain.

 

All maternal females survived until scheduled sacrifice and no adverse clinical signs of toxicity were observed. Maternal food consumption was statistically significantly reduced in the 12000 ppm group from Days 2-18 inclusive of pregnancy. There were no adverse effects on food consumption in the 300 or 1800 ppm test material group. In line with the reduction in food consumption in the high-dose females, administration of 12000 ppm test material was associated with a small but statistically significant reduction in bodyweight gain compared with the control group which was most marked at the start of the feeding period. Maternal macroscopic findings (post mortem) were considered not to be related to test material treatment. There was no effect at any dose on foetal weight, litter weight, gravid uterus weight, numbers of intra-uterine deaths or numbers of external abnormalities.

 

Examination of the feotuses revealed that the incidence of minor external and visceral defects was unaffected by treatment although two visceral variants were increased at the top two dose levels; kinked ureter being increased in the 1800 and 12000 ppm groups and slightly dilated ureter being increased in the 12000 ppm group. Overall, minor skeletal defects were increased in a dose-related manner at 1800 and 12000 ppm test material, while skeletal variants and pes score were increased at the top dose only. These findings indicate slightly poorer ossification at the 1800 and 12000 ppm dose levels. The reduced ossification and increase in the incidence of visceral variants are considered to be the result of slight foetotoxicity. There was no treatment-related effect on skeletal or visceral variants at 300 ppm test material.

 

Overall, under the conditions of the study, the NOAEL (maternal toxicity), based on reduction in body weight and food intake at 12000 ppm (1080 mg/kg), was determined to be 170 mg/kg bw/day while the NOEL (fetotoxicity), based on reduced ossification and increase in the incidence of visceral variants observed at the highest doses, was determined to be 28 mg/kg bw/day. These effects were not considered to be adverse.


Justification for selection of Effect on developmental toxicity: via oral route:
Only one study is available.

Justification for classification or non-classification

In accordance with with criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the material does not need to be classified for toxicity to reproduction.